This invention relates to pharmaceutically active composition extracted from Ferula hermonis and process for its preparation using supercritical carbon dioxide.
Ferula hermonis (known as Shirsh Zallouh) has recently received much attention due to its commercial value as an aphrodisiac, and as an herbal alternative to pharmaceutical drugs without the side effects. Shirsh Zallouh is the Arabic for xe2x80x9chairy rootxe2x80x9d, a perennial of the parsley family. In northern Lebanon, Shirsh Zallouh is called Hashishat al-Kattira, which means herb of abundance. Ferula hermonis is a small shrub, with pale pink flowers. It grows at more than 6000 feet on the high mountain areas of northern Lebanon, and on the biblical Mount Hermon in Southern Lebanon, at the joint borders of Syria and Israel.
Ferula hermonis roots are usually picked in the summer, after they mature and before the snow-season. There are six different species of the plant Ferula, one of them contained the poison that killed the ancient Greek Philosopher Socrates. Ferula hermonis belongs to the family of plants known as the Umbelliferae. This family contains plants like the carrot, fennel, Chinese angelica and hemlock. Hemlock, of course, is extremely poisonous and there are reports that naxc3xafve ferula hunters are endangering their lives by picking hemlock, which is almost identical in appearance to Ferula.
Ferula genus constitutes about 150 species, and these are widely distributed throughout the Mediterranean area to central Asia. Medicinal properties of Ferula plants include antispasmodic, carminative, digestive, expectorant, sedative, antihysteric, laxative, aphrodisiac, antiseptic, and analgesic. The genus ferula species are rich in the sesquiterpenes daucanes, humulanes, carotanes, himachalanes, and guaianes.
Ferula hermonis"" sexual potency was discovered by goat herders who noticed its strong sexual effects on their herds after eating this plant during the mating season. Middle East herbalists have used Ferula hermonis for centuries as a folk remedy to treat frigidity in women, and erectile and sexual dysfimction in men by increasing blood flow to sexual organs with dazzling results, and have reported renewed vigor, potency and energy. Ferula hermonis has been used by the elderly, in the Middle East for centuries to reinvigorate their sex lives.
In humans, the aphrodisiac property and safety of Ferula hermonis has been demonstrated in clinical trials involving more than 7,000 men with erectile dysfunction. The studies found that Ferula hermonis improved sexual function, increased libido and firmness of erections within few days. Ferula hermonis may work immediately for some, and may take few weeks for others. These trials revealed that 60 to 88 percent of men with erectile dysfunction experienced improved erections and increased desire within a few weeks after taking Ferula hermonis root; less than 4 percent experienced side effects such as headaches and flushing. The studies also found that Ferula hermonis may work almost immediately for some, and may take several weeks for others.
Because of the vasodilatation effects of Ferula hermonis, people with cardiovascular or neurological disorder should consult a heath care professional before taking this supplement. The common side effects associated with Ferula hermonis are headaches, flushing, and gastrointestinal tract.
Previous phytochemical investigations on Ferula hermonis revealed the presence of xcex1-bisabolol and the daucane sesquiterpenes: 8,9-epoxy jaeschkeanadiol benzoate (epoxy ferutinol benzoate, (3)), jaeschkeanadiol vanillate (ferutinol vanillate, (4)), jaeschkeanadiol p-hydroxybenzoate (ferutinin, (1)), jaeschkeanadiol benzoate (Teferdin, (2)), jaeschkeanadiol (ferutinol, (5)), 8,9-epoxy jaeschjeanadiol (12), 14-(4 hydroxybenzoyloxy) dauc-4,8-diene (6), and 14-(4xe2x80x2-hydroxy-3xe2x80x2-methoxy-benzoyloxy) dauc-4,8-diene (7).
Ferutinine (1) and Tenuferidine (11) have been shown to have estrogenic activity, and may contribute to its aphrodisiac activity. Zallouh root also contains naturally occurring vitamins (A, B 1, B2, B6, C, D, and E) and minerals (iron, magnesium, selenium, and zinc).
A recent study found that Ferutinin, Ferutidin, and Tenuferidin increase cation permeability of lipid bilayers and mitochondria in a dose-dependent manner (Biochemica et Biophysica Acta. 2001; 1512:98-110).
It has been suggested that some components of Panoferol (a mixture of terpenoids from Ferula) may increase sex hormone levels and calcification rates, suggesting that panoferol acts on calcium homestasis. This suggestion was confirmed by the discovery that one of the panoferol mixture (Ferutinin) possesses Ca2+ionophoric properties (Ignatkov V I, Ahmedhodzjaeva H T, Babichev V. effects of Tefestrol on the secretion of luteinizing hormone from the hypophysis. Farmakol Toksikol 1990; 53:37-38).
Ferutinin at concentration range 1 to 50 micromolar increased the permeability of thymocytes, mitochondria, sarcoplasmic reticulum, liposomes and bilayer lipid membranes for Ca2+(Zamaraeva M V, Hagelgans A I, Abramov A Y, Ternovsky V I, Merzlyak P G, Tashmukhamedov B A, Saidkhodzjaev A. lonophoric properties of ferutinin. Cell calcium 1997; 22:235-241) Compounds Salpha-(3-methoxy-4-hydroxybenzoic acid) ester of jaeschjeanadiol, U and 5alpha-(p-hydroxybenzoic acid) ester ofjaeschjeanadiolxe2x80x2were found to prevent pregnancy in adult female rats when administered orally on 1 to 7 post coitum. Compound Salpha-(3-methoxy-4-hydroxybenzoic acid) ester ofjaeschjeanadiol was found to be more potent than 5alpha-(p-hydroxybenzoic acid) ester ofjaeschjeanadiol since it prevented pregnancy in rats after a single oral administration of 5 mg per kg dose on the first day of post coitum. These two compounds also exhibited potent estrogenic activity. In an in-vitro assay to measure relative binding of these compounds to immature rat uterine cytosol estrogen receptors, these compounds exhibited relative binding affinity of 0.01 percent and 5.75 percent of 17 beta-estrodiol, respectively, for immature rat uterine cytosol estrogen receptors (Singh M M, Agnihotri A, Garg S N, Agarwal S K, Gupta D N, Keshri G, Kamboj V P. Antifertility and hormonal properties of certain carotane sesquiterpenes of Ferula jaeschkeana. Planta Medica 1988; 492).
In a recent study, the oil extracted from Ferula harmonis was found to enhance sexual activities as assessed by penile erection index in a dose dependent manner in male rats. The effective dose (12.03 mg/kg) was 880 times less than the lethal dose LD(50) (10.6 g/kg). This study also found that the Ferula harmonis extract becomes toxic if it is used for a long period of time (El-Thaher T S, Matalka K Z, Taha H A, Badwan A A. Ferula harmonis xe2x80x98zallouhxe2x80x99and enhancing erectile function in rats: efficacy and toxicity study. Int J Impot Res 2001; 13:247-251).
The effects of Ferula hermonis extract on social aggression, fertility and some physiological parameters were examined in prepubertal male mice. Ingestion of 3 mg/kg of aqueous extract of Ferula hermonis for six weeks inhibited social aggression, and a significant reduction of their fertility. This treatment caused a significant decrease in the number of pregnant females, number of implantations and viable fetuses in females impregnated by males that ingested this extract (Khleifat K, Homady M H, Tarawneh K A, Shakhanbeh J. Effect of Ferula hermonis extract on social aggression, fertility and some physiological. parameters in prepubertal male mice. Endocr J 2001; 48:473).
The traditional way to take this herb is to slit the root and wait for the resin to ooze out. This process is now replaced by the more conventional organic solvent extraction procedure. Lebanese pharmacists cut up the root and make an extract by a hot alcohol extraction and distillation. It is recommended that men take 50 drops (two and half ml) of an alcohol (alcohol concentration of over ninety percent) extract of Zallouh root at night and again first thing in the morning. This regimen should be followed for at least four weeks. For women suffering from menstrual/menopausal complaints, 50 drops three times daily, after meals. The roots can also be soaked in wine or ground into powder and then taken in capsules or mixed with tea. In Syria, the powder is mixed with honey.
The suggested dose ranges from two to eight grams of Zallouh root taken as a tea. Some studies also indicated that a significant number of males experienced increased number of erections by taking Zallouh one to three hours prior to the anticipated sexual encounter.
The present invention overcomes the problem of using large effective dosage of Ferula hermonis, and provides a pharmaceutically active composition of Ferula hermonis free of solvent residue useful for small effective dosage and useful for formulation with other ingredients, and for formulation useful in different forms including soft and hard gel capsules and tablets, and clinical studies. The pharmaceutically active composition of Ferula hermonis free of solvent is obtained by supercritical fluid extraction.
The use of supercritical fluids for the extraction of chemical compounds from plants has received increasing attention because of the potential to dramatically reduce the time required for sample extraction as well as eliminating the need for large volumes of liquid solvents. Supercritical extraction is a technique in which gases are compressed under supercritical conditions to form a fluid, which is then used to remove chemicals from a matrix. Supercritical fluids have good extracting power because of their density, which can be controlled by changes in pressure or temperature, and to low viscosity, high diffusivity and low surface tension, which enhance mass transfer inside a solid matrix.
Carbon dioxide has a manageable critical point (i.e., critical pressure of 73 bar and critical temperature of 31 .degree. C.). At temperatures up to 31.06 degrees C. carbon dioxide can be liquefied by raising the pressure and this liquid exerts appreciable solvent power to dissolve natural oils and quite a wide range of non-polar or slightly polar materials. Supercritical carbon dioxide (CO2) has been the most frequently used solvent in supercritical fluid extraction, being non-toxic, non-flammable, inexpensive and easily separated from extracts. Furthermore, the low critical temperature (31.06 degrees C.) of CO2 allows extractions of heat sensitive compounds without degradation. As a result of these advantages, supercritical CO2 fluid extraction has received increased attention as an alternative to conventional separation methods (see, e.g., Kirk-Othmer Encyclopedia of Chemical Technology, pages 872-893, Supplement volume, third edition, 1984, John Wiley and Sons, New York). Supercritical fluids have found myriad uses, including extracting oils, flavors, fragrances, and other materials from foods (Dziezak J.D., Innovative separation process finding its way into the food industry. Food Technology 1986; 40:66-9); in pharmaceutical industries (Bruno J, Castro C. A. N., Hamel, J. F. P., Palavra A.M.F. Supercritical fluid extraction of biological products. In Recovery processesfor biological materials, eds Kennedy J. F. and Cabral J. M. S., John Wiley and Sons, Chichester 1993, pp.303-54); and for the extraction of natural products (Extraction of natural products using near-critical solvents. Edited by King M. B. and Bott T. R. Published by Chapman and Hall, 1993; and Rui L. Mendes et al. Applications of supercritical carbon dioxide extraction to microalgae and plants. J Chem Tech Biotechnol 1995; 62:53-59).
Some of the food applications include de-caffeination of coffee and tea, hops, oil recovery and extraction of spices (Rizvi S. S. H., Daniels J.A., Benado A. L. and Zollweg, J.A., Supercritical fluid extraction: operating principles and food applications. Food Technology 1986; 40:57-64). Pharmaceutical applications include extraction of steroids (Larson K. A., and King M. L., Evaluation of supercritical fluid extraction in the pharmaceutical industry. Biotech. Prog 1986; 2:73-82) and chemotherapeutic alkaloids (Schaeffer S. T., Zalkow L. H., and Teja A. S., Extraction and isolation of chemotherapeutic pyrrolizidine alkaloids from plant substrates. ACS Symposium Series 1989; 406:416-33).
This technique has been used on an industrial scale for over two decades for the extraction of flavor principles from hops and other herbs. It has the advantage over extraction with conventional organic solvents that the extraction medium is readily and completely removed by allowing the liquid carbon dioxide to vaporize into the atmosphere. Thus, problems of waste solvent disposal and trace solvent contamination of finished product are eliminated. The uses of supercritical carbon dioxide for the preparation of plant extracts have been described in a number of U.S. patents.
U.S. Pat. No. 4,104, 409 described a process for extraction of the resin and essential oils of hops utilizing supercritical carbon dioxide fluid.
U.S. Pat. No. 6,180,105 disclosed a method of preparation of an artemisinin extract comprising the steps of extraction of Artemisia annua with liquid carbon dioxide and allowing the carbon dioxide to evaporate from the resultant mixture.
U.S. Pat. No. 6,319,524 disclosed a method of extracting saw palmetto berries by contacting ground saw palmetto berries with carbon dioxide at a pressure of at least 500 bar, and at a temperature of less than about 80 degrees C.
U.S. Pat. No. 6,117,431 described a method involving supercritical carbon dioxide to produce a purified extract from ginkgo biloba leaves.
U.S. Pat. No. 5,466,451 disclosed a process for extraction of pharmaceutically active compositions from the plant Tanacetum parthenium (Feverfew).
U.S. Pat. No. 5,591,343 disclosed a process for extraction of carotenoids from bacterial cells comprising the step of bringing the bacterial cells into contact with supercritical fluid so as to extract the carotenoids from the cells.
U.S. Pat. No. 6,111,108 described a method for extraction of biologically active components from the plant Camptotheca using supercritical carbon dioxide.
The present invention overcomes the problem of using large effective dosage of Ferula hermonis, and provides a pharmaceutically effective composition of Ferula hermonis free of solvent residue useful for small effective dosage and useful for formulation with other ingredients such as, but not limited to, sexual enahnacement, diabetic, weight management, hair, and tonic; useful formulation in different forms including soft and hard gel capsules and tablets; and clinical studies. The pharmaceutically active composition of Ferula hermonis free of solvent is obtained by supercritical fluid extraction.
It is an object of the invention to provide a pharmaceutically active composition from Ferula hermonis, said composition obtained by a process comprising fmely milling Ferula hermonis, and extracting the fmely milled Ferula hermonis with carbon dioxide in the supercritical state at a temperature from 31.degree to 80.degree. C. and under a pressure from 150 to 400 bar. According to a further aspect of the invention, the objects are achieved by providing a process for extracting a pharmaceutically active composition from Ferula hermonis said process comprising extracting finely milled Ferula hermonis with carbon dioxide in the supercritical state at a temperature from 31.degree to 80.degree. C. and under pressure from 150 to 400 bar.
It is also an object of the invention to provide a pharmaceutically active composition from Ferula hermonis containing terpenoid -like compounds as ingredients, which predominate in terms of amount.
It is also an object of the invention to provide a pharmaceutically active composition from Ferula hermonis for use in the preparation of pharmaceutical formulas, soft-drink, beverages, infusion, tablets, capsules, soft-gel capsules and dietary supplements.
Extracting is accomplished by flowing supercritical carbon dioxide through the powdered Ferula hermonis at an extraction pressure of at least about 150 bar. Those skilled in the art will recognize that a xe2x80x9cbarxe2x80x9d is a unit of pressure substantially equivalent to one atmosphere, or 10. sup.5 newton/m.sup.2.Following extraction, the pharmaceutically active composition is separated from the carbon dioxide by decreasing the pressure to a predetermined separation pressure lower than the extraction pressure, and at a temperature sufficient to prevent the carbon dioxide from solidifying.
These objectives of the present invention will be more readily appreciated and understood from a consideration of the following detailed description.